Field of the Invention
The present invention relates to a biomolecule extraction device and biomolecule extraction method. More specifically, it relates to a biomolecule extraction device and biomolecule extraction method that can enhance a user's convenience by simplifying the process of extracting a biomolecule, such as a protein or a nucleic acid, etc., from a biomass sample, such as tissue and cell samples, with a lyses buffer, and saving time spent. As the ratio of the surface area of the sample to the amount of lysis buffer introduced is maximized, the concentration of biomolecule extraction is increased and the amount of buffer used is minimized. The present invention can also smoothly perform efficient testing without extra device or equipment by allowing constant discharge of the biomass lysate containing the extracted biomass.
Description of Related Art
Cell lysis refers to a phenomenon where a cell membrane is ruptured and cell contents (cytoplasma) are exposed as the cell dissolves. Such cell lysis is a primary process for cell analysis and protein purification, and is widely used not only to extract/separate protein, but also to separate nucleic acid, such as DNA (deoxyribonucleic acid) or RNA (ribonucleic acid) prior to an amplification process, such as the PCR (polymerase chain reaction), used in molecular biology and molecular diagnostics, etc.
Cell lysis methods for cell disruption largely include optical, acoustic, electrical and mechanical methods. Mostly, the methods are carried out in a form applying external force and stress in various mechanical and physical manners based on a lysis buffer.
Optical cell lysis is a method for destroying cells by irradiating laser micropulse on the target cells to form cavitation bubbles and destroying cells as the cavitation bubbles expand. Optical cell lysis has disadvantages in that there is a possibility for the cell and protein to be degenerated due to the heat abruptly generated by applying a laser inside a specific cell or at a nearby location, and that a separate device for generating laser micropulses must be used should be added.
Acoustic cell lysis is a method for destroying cells by introducing a cell solution or a suspension inside a chamber located in an ultrasonic water tank and applying ultrasonic waves. It is difficult to obtain consistent results for cell destruction using ultrasonic waves because it is difficult to form uniform energy distribution of ultrasound waves, it takes a lot of time for cell destruction, and the cell destruction using ultrasound waves may cause protein destruction or deformation due to the heat generated.
Electrical cell lysis is a method destroying cells by applying an electric field to the cells to generate potential difference in cell membrane. It is similar in a way to other cell lysis methods such as the freezing-thawing method, heating method, osmotic pressure impact method, in terms of applying impact to cell walls. However, these methods have problems that protein in the cells may be damaged due to the thermal impact applied to the cells.
Mechanical cell lysis uses presses, bead mills, etc. To be specific, presses perform cell disruption by filling an empty cylindrical body made of stainless steel often used in laboratory scale with cell paste, and extracting the cells to atmospheric pressure through a needle valve on the bottom of the cylinder under high pressure.
High-speed bead mills comprise a grinding chamber filled with small glass or iron beads (20˜50 units), and grind cells with high shearing force and impact force by rotating a circular disk or impeller attached to a driving shaft by a motor and stirring the beads.
Such mechanical cell lysis has problems that it is difficult to apply the mechanical cell lysis to a small amount of samples, and expensive equipment, large space, multi-step process and long processing time are required.
Meanwhile, a homogenizer performs cell lysis and disruption by having a user rotate a stick while filling an E-tube (Eppendorf-tube) or falcon tubes of various capacities, etc. with a lysis buffer. In this regard, there may be problems that in order to immerse a tape or disk for obtaining samples, a relatively large amount of lysis buffer is required and the sample may splash out or the buffer may overflow while rotating the stick.
In addition, there are problems that in case of using the E-tube (Eppendorf-tube) or Petri dish itself, in order to apply the lysis buffer to a hydrophobic sample, a relatively large amount of buffer is required and additional work such as several times of pipetting, etc. using tools such as a pipette is required.
Meanwhile, cell lysates according to cell lysis are widely used for special protein detection tests (western blotting) or immune precipitation, etc., and in case of extracting nucleic acid (DNA, RNA), they are applied to molecular diagnostics and gene analysis, etc. using PCR or sequencing. The above processes are performed by detecting the special protein itself or testing interaction between molecules.
Here, for cell lysis, it is preferable to have a sufficient amount of biomolecule (protein or nucleic acid, etc.) product extracted from biomass, high purification concentration, and no loss or deformation of the extract. For doing so, an expensive protease inhibitor, etc. is used. Thus, since cell lysis of good quality for biomass sample should be performed fast and analyzed immediately, it would be necessary to simplify the process of extracting biomolecule and save cost and time spent therefor.
However, as mentioned above, the conventional biomolecule extraction devices and methods thereof through cell lysis and disruption required an extra device or equipment such as a centrifugal separator and pipette, etc. while performing each steps. Plus, a complex process associated with the system had to be performed, thereby requiring a lot of space, cost and time.
Also, in case of using a tape or disk for obtaining samples, there are problems that a lysis buffer where the sample can be immerged is required in order to maximize the contact surface of the sample and that due to the decrease in extraction concentration by applying a large amount of lysis buffer, a large amount of samples, i.e., biomass needs to be collected and additional work of chopping or dissociating the sample is required.
This still leaves the problem of processing the residue which may contaminate the environment and affect human safety, after extracting biomolecule such as protein or nucleic acid from the sample.
Thus, the necessity of a biomolecule extraction device has been raised where user's convenience can be enhanced by saving time and reducing space in need, resulted from simplifying the process of extracting biomolecule from biomass, and where consistent test results can be expected by minimizing the damage of the extracted biomolecule and extracting at a relatively high concentration while applying only a small amount of samples.
Further, the biomolecule extraction device can not only increase the concentration of biomolecule extracted and reduce the amount of buffer used by minimizing dead space thereby maximizing the ratio of the surface area of the sample to the amount of lysis buffer introduced, but can also efficiently and smoothly perform testing by allowing constant biomass lysate containing the extracted biomass.